School of Basic Medical Sciences, Guangdong Pharmaceutical University, Guangzhou, 510006, China; Guangdong Key Laboratory of Bioactive Drug Research, Guangzhou, 510006, China.
School of Basic Medical Sciences, Guangdong Pharmaceutical University, Guangzhou, 510006, China.
Environ Res. 2024 Mar 1;244:117676. doi: 10.1016/j.envres.2023.117676. Epub 2023 Nov 21.
The soil environment for straw return is a rich and valuable library containing many microorganisms and proteins. In this study, we aimed to screen a high-quality β-glucosidase (BGL) from the soil metagenomic library and to overcome the limitation of the low extraction rate of resveratrol in Polygonum cuspidatum. This includes the construction of a soil metagenomic library, screening of BGL, bioinformatics analysis, cloning, expression, immobilization, enzymatic property analysis, and application for the transformation of polydatin. The results showed that the soil metagenomic library of straw return was successfully constructed, and a novel BGL was screened. The identified 1356 bp long BGL belonged to the glycoside hydrolase 1 (GH1) family and was named Bgl1356. After successful cloning and expression of Bgl1356, it was immobilized using chitosan. The optimum temperature of immobilized Bgl1356 was 50 °C, and the pH was 5. It exhibited good tolerance for various metal ions (CO, Ni, Cu, Mn, Na, Ca, and Ag) and organic solvents (DMSO, Triton-X-10, and ethanol). Enzymatic kinetics assays showed that Bgl1356 had good affinity for the substrate, and the specific enzyme activity was 234.03 U/mg. The conversion rate of polydatin by immobilized Bgl1356 was 95.70 ± 1.08%, facilitating the production of high amounts of resveratrol. Thus, this paper reports a novel temperature-, organic solvent-, and metal ion-tolerant BGL that has good application prospects in the pharmaceutical industry.
秸秆还田土壤环境是一个富含多种微生物和蛋白质的宝贵文库。本研究旨在从土壤宏基因组文库中筛选出高质量的β-葡萄糖苷酶（BGL），并克服虎杖中白藜芦醇提取率低的限制。这包括构建土壤宏基因组文库、筛选 BGL、生物信息学分析、克隆、表达、固定化、酶学性质分析以及在虎杖苷转化中的应用。结果表明，成功构建了秸秆还田土壤宏基因组文库，并筛选出一种新型 BGL。鉴定出的 1356bp 长 BGL 属于糖苷水解酶 1（GH1）家族，命名为 Bgl1356。成功克隆和表达 Bgl1356 后，用壳聚糖对其进行固定化。固定化 Bgl1356 的最适温度为 50°C，最适 pH 为 5。它对各种金属离子（CO、Ni、Cu、Mn、Na、Ca 和 Ag）和有机溶剂（DMSO、Triton-X-10 和乙醇）具有良好的耐受性。酶动力学分析表明，Bgl1356 对底物具有良好的亲和力，比酶活为 234.03 U/mg。固定化 Bgl1356 对虎杖苷的转化率为 95.70±1.08%，有利于生产大量白藜芦醇。因此，本研究报道了一种新型耐温、耐有机溶剂和耐金属离子的 BGL，在制药工业中有很好的应用前景。
